Littérature scientifique sur le sujet « Marcatori biologici »

Il guadagno diagnostico offerto dalla RM nella diagnosi dei meningiomi intra-cranici risiede nella precisa localizzazione della lesione all'interno del cranio e nella definizione dei suoi rapporti con le adiacenti strutture, in particolare arterie e vene. Le caratteristiche morfo-strutturali possono essere comparate con gli aspetti istologici ed operatori in modo da identificare possibili caratteri clinico-radiologici associati ad un comportamento biologico ed evolutivo diverso da quello usualmente benigno dei meningiomi non aggressivi (MNA). 64 meningiomi intra-cranici sono stati valutati retrospettivamente, tutti sottoposti ad esame RM ed operati. In 16/64 pazienti (25%) è stato asportato un meningioma «aggressivo» (MA), caratterizzato cioè istologicamente da pleomorfismo ed alta cellularità, prominenza di nucleoli, frequenti mitosi e foci necrotici, e macroscopicamente da invasione locale della dura, dell'osso e dei tessuti epicranici. MA sono stati riscontrati più frequentemente nei maschi (9/16) di età avanzata con localizzazione alla convessità (12/16). All'esame RM presentavano accentuazione disomogenea (7/14), e marcatamente estesa alla dura adiacente alla base d'impianto («string sign») (10/16), scarsa demarcazione dal tessuto cerebrale circostante (6/14), edema marcato (11/16) e presenza di numerosi foci necrotici all'interno della lesione (7/16). La valutazione statistica ha permesso di individuare quei parametri (accentuazione disomogenea, necrosi, «string sign» ed edema marcato) la cui presenza o meno influisce probabilisticamente sulla diagnosi di MA. In particolare, quando nessuno dei parametri è presente, si tratta di un MNA; quando, al contrario, 3 o 4 parametri sono presenti, la lesione in esame rappresenta un MA; quando solo 1 o 2 parametri sono presenti, è impossibile stabilire con certezza la natura del meningioma.

Background:Clinical practice with patients suffering from chronic diseases highlights the presence of psychological symptoms of discomfort fed by biological and non-biological mechanisms linked to disease and treatment. In rheumatic diseases, literature detects the presence of anxious symptoms and depressed mood of clinical and sub-clinical importance with a multifactorial genesis1.Objectives:To detect the impact on the state of health of anxious symptoms and depressed mood in a population suffering from RA and other rheumatic diseases in order to implement the effectiveness of psychological intervention through the selection of patients who present critical levels of discomfort.Methods:Patients afferent to the Rheumatology outpatient clinic of Mauriziano Hospital have been screened from May 2018 to July 2018 with two self-administered questionnaires: HADS-A and HADS-D (Hospital Anxiety and Depression Scale), specifically developed for the evaluation of anxious and depressive symptoms in medical pathologies, and HAQ (Health Assessment Questionnaire) to explore functional disability. Data about rheumatic diagnosis and socio-demographic characteristics were also collected. Data were analyzed with descriptive statistics; the Student Test and the ANOVA test were used to evaluate prevalence and to compare the presentation of symptoms in the different diseases and the Pearson correlation coefficient was used to evaluate the relationship between symptoms and disability.Results:A total of 427 subjects were screened (317 females and 110 males), aged between 19 and 90 years (mean 60 ± 14 yrs). 156 subjects (36.5%) had a diagnosis of RA, 76 (17.8%) of psoriatic arthritis, 42 (9.8%) of ankylosing spondylitis, 14 (3.3%) of systemic lupus erythematosus and 139 (32.6%) of other rheumatic diseases (including Sjogren, osteoarthritis, fibromyalgia).A high prevalence of anxious symptoms and depressed mood has been found and the number of subjects reporting scores indicating a clinically relevant uncomfortable situation (HADS ≥ 11) was also relevant (Table 1); an increased prevalence in female patients was observed. There were no differences in the presentation of symptoms between RA and the other included pathologies (Table 2).Table 1.Prevalence of anxiety and depression according to the HADS questionnaire in rheumatic diseasesMeanSDHADS-A7.564.63HADS-D7.124.59HADS-A ScoreN%0-722452.47-108419.711-2111927.9HADS-D ScoreN%0-723154.17-109221.511-2110424.4Table 2.Comparison between RA and other rheumatic diseases in anxiety and depression symptoms presentation (ANOVA test).NMeanSDSECIHADS-ARA1562.345.200.411.52PsA762.304.470.511.28AS421.513.190.490.51SLE141.773.741.00-0.38other1392.465.080.431.61HADS-DRA1561.743.510.281.19PsA762.034.210.481.07AS420.690.540.080.52SLE140.930.680.180.54other1391.683.790.321.04There was a positive and significant correlation between anxious symptoms or depressed mood and functional disability (0.49 and 0.60 respectively, p<0,01).Conclusion:The results show a significant presence of uncomfortable situations that could evolve in a psychopathological sense. The discomfort expressed through anxious and depressive symptoms is related to the level of functional disability. Recognizing the presence of psychological distress allows to orient the treatment plan and facilitate the patient’s adaptation to the disease condition.References:[1]Geenen R. et al. Best Pract Res Clin Rheumatol. 2012;26(3):305-19.Disclosure of Interests:Gloria Crepaldi Consultant of: Advisory board for Sanofi and Celgene, Speakers bureau: BMS, MSD, Mariarosaria Voci: None declared, Marta Saracco: None declared, Antonella Laezza: None declared, Paolo Santino: None declared, Maddalena Marcato: None declared, Guido Rovera: None declared, Claudia Lomater Consultant of: Advisory board for Sanofi, Novartis, Abbvie

BackgroundAdoptive cell therapies with T lymphocytes expressing engineered T cell receptors (TCRs) are one of the most promising approaches to cancer therapy.1 However, the experimentally driven development of novel TCR therapies is limited by the enormous biological variability of peptide:Human Leukocyte Antigen:TCR (pHLA:TCR) complexes. The in silico methods hold the promise to streamline the discovery of novel TCR therapies by reducing costs and time of laboratory research. In particular, the prediction of TCR binding to a target antigen, as well as the prediction of TCR off-target toxicity2 can provide useful insights supporting the development of safe therapies. We aimed at the development of an experimentally validated AI model of pHLA:TCR binding that will help to prioritize and reduce the number of in vitro assays necessary to discover novel TCRs for cancer therapies.MethodsThe limiting factor of successful pHLA:TCR binding modeling is data availability and completeness of TCR characterization. To address this issue, we are building an oncological pHLA:TCR database with paired alpha and beta chain TCR sequences. We are collecting and sequencing tumor and normal samples from 100 cancer patients, as part of an observational clinical trial. Those data are then screened with the Ardigen's ArdImmune Vax platform3 4 to select immunogenic epitopes. T cells that bind those epitopes are subsequently sorted and used to generate TCR sequencing data at single-cell resolution. We use data-driven and simulation-based models to extract insights about the dynamics of a pHLA:TCR system to predict the binding probability and explain the inference made by the model.ResultsWe optimized our data collection pipeline for the cost-efficient acquisition of a large oncological pHLA:TCR dataset. These data will enable us to build efficient models to streamline the development of TCR therapies against cancer.We benchmarked our modeling approach for pHLA:TCR binding against existing solutions5–7 on publicly available data. We also show how focus on model explainability facilitates the detection of model inconsistency of uncertain predictions by expert inspection. Our toxicity assessment solution2 extends the applicability of our system to the prediction of TCR safety profile.ConclusionsThe presented work shows perspectives and limitations of AI-aided TCR therapy development. We present results for our pHLA:TCR binding model, a TCR-toxicity-screening solution, and the study design of our observational clinical trial. Our growing database of pHLA:TCR interactions will enable us to develop highly predictive pHLA:TCR binding models, in particular for oncological targets.AcknowledgementsWe acknowledge funding through the project “Creating an innovative AI-based (Artificial Intelligence) IN SILICO TECHNOLOGY TCRact to launch a NEW SERVICE for designing and optimizing T-cell receptors (TCR) for use in cancer immunotherapies” cofunded by European Regional Development Fund (ERDF) as part of Smart Growth Operational Programme 2014–2020.ReferencesFarkona S, Diamandis EP, Blasutig IM. Cancer immunotherapy: the beginning of the end of cancer? BMC Med 2016;14:73. PMCID: PMC4858828.Murcia Pienkowski VA, Mazzocco G, Niemiec I, Sanecka-Duin A, Krol P, Myronov O, Skoczylas P, Kaczmarczyk J, Blum A. Off-target toxicity prediction in cellular cancer immunotherapies [Internet]. Cytotherapy. 2021;S96. Available from: http://dx.doi.org/10.1016/s1465324921004229.Stepniak P, Mazzocco G, Myronov A, Niemiec I, Gruba K, Skoczylas P, Sanecka-Duin A, Drwal M, Kaczmarczyk J. AI-augmented design of effective therapeutic cancer vaccines and adoptive cell therapies. Journal For Immunotherapy Of Cancer. Bmc Campus, 4 Crinan St, London N1 9xw, England; 2019.Mazzocco G, Niemiec I, Myronov A, Skoczylas P, Kaczmarczyk J, Sanecka-Duin A, Gruba K, Król P, Drwal M, Szczepanik M, Pyrc K, Stȩpniak P. AI aided design of epitope-based vaccine for the induction of cellular immune responses against SARS-CoV-2. Front Genet. 2021;12:602196. PMCID: PMC8027494.Weber A, Born J, Rodriguez Martínez M. TITAN: T-cell receptor specificity prediction with bimodal attention networks. Bioinformatics. 2021;37(Suppl_1):i237–i244. PMCID: PMC8275323.Springer I, Besser H, Tickotsky-Moskovitz N, Dvorkin S, Louzoun Y. Prediction of specific TCR-peptide binding from large dictionaries of TCR-Peptide Pairs. Front Immunol 2020;11:1803. PMCID: PMC7477042.Jurtz VI, Jessen LE, Bentzen AK, Jespersen MC, Mahajan S, Vita R, Jensen KK, Marcatili P, Hadrup SR, Peters B, Nielsen M. NetTCR: sequence-based prediction of TCR binding to peptide-MHC complexes using convolutional neural networks [Internet]. Available from: http://dx.doi.org/10.1101/433706

Recent reports showed that early-interim PET-scan is the only tool predicting treatment outcome in advanced-stage classical Hodgkin lymphoma (asCHL). We evaluated the prognostic impact of a series of immunohistochemical markers, mentioned in literature as prognostic factors, on tissue microarrays assembled from biopsies of 220 patients: STAT1, SAP, TOP2A, PCNA and CD20, both in neoplastic (HRSC) and microenvironment cells (MC); RRM2, MAD2, CDC2, BCL2, P53, BCL11A and EBER in HRSC; ALDH1A1, TIA-1, granzyme B, perforin, FOXP3, and PD-1 in MC. All patients had been treated with standard ABVD ± Rx therapy. Interim-PET after 2 ABVD courses was evaluated according to the criteria indicated by Gallamini in his study (Journal of Clinical Oncology, 2007). The survival analysis has been performed in a subset of 138 patients whose complete clinical information were available: the mean age was 33.3 years (14-79), the stage III-IVB in 98 and IIB in 40, and the mean follow-up 38.1 months (7.6-71.9). Histopathology review showed: NS-I 75, NS-II 22, MC 20, DL 3, and CHL/nos 18 cases. Interim-PET was positive in 30 patients, while treatment failure was recorded in 32. In univariate analysis the factors related to treatment outcome were BCL2 on HRSC (cut-off value 50%), STAT1/SAP on MC, and PET (Log-rank 6.9, 7.9 and 93.9 respectively). The combined expression of STAT1 and SAP was scored in three levels depending on the architectural pattern: score 0 for expression of both with a diffuse/rosetting pattern; score 1 for discordant combination of diffuse/rosetting and scattered patterns; score 2 for both markers with a scattered pattern; the 3y-PFS were 87.4%, 69.9% and 61.9% respectively. In multivariate analysis PET, BCL2 and STAT1/SAP remained significant (HR: 24.8, 4.6, 7.5 and 5.6, respectively; p<.01). The proposed model is able to predict treatment response in AsCHL, even if with a lower efficacy than PET. However, unlike PET, it can be applied upfront therapy.

Recent reports showed that early-interim PET-scan is the only tool predicting treatment outcome in advanced-stage classical Hodgkin lymphoma (asCHL). We evaluated the prognostic impact of a series of immunohistochemical markers, mentioned in literature as prognostic factors, on tissue microarrays assembled from biopsies of 220 patients: STAT1, SAP, TOP2A, PCNA and CD20, both in neoplastic (HRSC) and microenvironment cells (MC); RRM2, MAD2, CDC2, BCL2, P53, BCL11A and EBER in HRSC; ALDH1A1, TIA-1, granzyme B, perforin, FOXP3, and PD-1 in MC. All patients had been treated with standard ABVD ± Rx therapy. Interim-PET after 2 ABVD courses was evaluated according to the criteria indicated by Gallamini in his study (Journal of Clinical Oncology, 2007). The survival analysis has been performed in a subset of 138 patients whose complete clinical information were available: the mean age was 33.3 years (14-79), the stage III-IVB in 98 and IIB in 40, and the mean follow-up 38.1 months (7.6-71.9). Histopathology review showed: NS-I 75, NS-II 22, MC 20, DL 3, and CHL/nos 18 cases. Interim-PET was positive in 30 patients, while treatment failure was recorded in 32. In univariate analysis the factors related to treatment outcome were BCL2 on HRSC (cut-off value 50%), STAT1/SAP on MC, and PET (Log-rank 6.9, 7.9 and 93.9 respectively). The combined expression of STAT1 and SAP was scored in three levels depending on the architectural pattern: score 0 for expression of both with a diffuse/rosetting pattern; score 1 for discordant combination of diffuse/rosetting and scattered patterns; score 2 for both markers with a scattered pattern; the 3y-PFS were 87.4%, 69.9% and 61.9% respectively. In multivariate analysis PET, BCL2 and STAT1/SAP remained significant (HR: 24.8, 4.6, 7.5 and 5.6, respectively; p<.01). The proposed model is able to predict treatment response in AsCHL, even if with a lower efficacy than PET. However, unlike PET, it can be applied upfront therapy.

Introduzione. L’adenocarcinoma duttale (PDAC) rappresenta circa l’85% delle neoplasie maligne pancreatiche. È un tipo di neoplasia molto aggressiva e ha prognosi infausta. Il PDAC è una malattia con una elevata mortalità, spesso diagnosticata in uno stadio avanzato per il quale esistono poche e inefficaci terapie. L’alta incidenza di ricadute locali unita alla precoce metastatizzazione sono le caratteristiche cliniche più tipiche di questo tumore. Inoltre la nota resistenza del tumore alla chemio e alla radioterapia limita l’efficacia di questi approcci terapeutici. Scopo. Identificare e validare nuovi marcatori biologici associati a caratteristiche di aggressività dell’adenocarcinoma pancreatico al fine di utilizzarli per comprendere proprietà biologiche del tumore stesso o in clinica per una corretta valutazione prognostica. Metodi e risultati. Abbiamo studiato n=17 linee cellulari umane immortalizzate di PDAC per alcune caratteristiche di aggressività cellulare: per la clonogenicità e la chemioresistenza alla gemcitabina in vitro e, in vivo, per la capacità di crescita in topi immunocompromessi (CD1-nude). Tutte le 17 linee cellulari sono state caratterizzate per l’espressione di classi di marcatori molecolari: recettori delle chemochine (CCR1-CCR10; CXCR1-CXCR6; CX3CR1; XCR1) e putativi marcatori staminali tumorali (ESA+CD24+CD44+; CD133+, CXCR4+) mediante citometria a flusso, secrezione di fattori solubili (n=48) tramite la tecnologia luminex e l’espressione di geni (n=11) coinvolti nello sviluppo pancreatico con Real Time PCR. Usando l’analisi statistica inter-linea (regressione lineare o di cox) abbiamo cercato una correlazione tra i fenotipi biologici e le caratteristiche di malignità cellulare individuando nuovi marcatori. Questi marcatori sono stati validati su tessuti tumorali primari in casistiche di pazienti affetti da PDAC: l’espressione del marcatore identificato è stata correlata con l’esisto clinico della neoplasia. In una prima analisi inter-linea n=35 fattori sono risultati statisticamente associati ad una o più caratteristiche di aggressività. È seguita una classificazione per priorità che, avvalendosi della sola correlazione con la tumorigenicità in vivo, ha ridotto a n=20 i fattori di rischio da validare. Abbiamo quindi approfondito lo studio su 4 marcatori molecolari di sviluppo pancreatico (ISL1, PDX1, PAX6, KRT19), sui fenotipi staminali e su 2 recettori delle chemochine (CCR5, CXCR3). L’espressione genica dell’mRNA di ISL1, PDX1, PAX6 e KRT19 è stata valutata in sezioni criostatiche di n=42 resezioni chirurgiche di pazienti affetti da PDAC. Non sono emerse correlazioni significative tra l’espressione di questi fattori e la sopravvivenza globale. Tuttavia alti livelli dell’mRNA di KRT19 predicono una progressione più precoce e di tipo metastatico. Più elevati livelli di PDX1 e PAX6 sono associati con una più alta probabilità di recidiva locale. Inoltre combinando i marcatori è stato individuato un fenotipo più aggressivo correlato con una minor sopravvivenza: si tratta dei pazienti che esprimono ad elevati livelli sia PDX1 che KRT19. La nostra strategia di screening ha mostrato essere fattori di rischio per lo sviluppo tumorale nel topo, non i classici fenotipi staminali descritti in letteratura (ESA+/CD24+/CD44+, CD133+, CD133+/CXCR4+) ma la combinazione ESA+/CD24-/CD44+ e la sola espressione di CXCR4 ed ESA: tuttavia la validazione clinica, condotta su una coorte di 39 pazienti affetti da adenocarcinoma duttale, non ha confermato che questi marcatori, né i classici già descritti, correlino in maniera statisticamente significativa con la sopravvivenza o con la progressione nel tempo e nemmeno con il sito di recidiva. Il fenotipo ESA+/CD24+/CD44- è invece risultato un fattore di rischio prognostico indipendente sia per la sopravvivenza (HR=4,166 p=0,001) che per la progressione (HR=2,208 p=0,019). CCR5 e CXCR3 sono risultati espressi su tessuti tumorali processati a fresco (n=6) ed analizzati in citometria a flusso a fronte di una negatività del pancreas di donatori d’organo (n=10). Essi sono espressi rispettivamente nell’11,9% e nel 17,6% delle cellule CA19.9+ del tumore, mentre solo nello 0,4% e il 0,34% nel tessuto sano. L’aumentata espressione di CCR5 e CXCR3 sembra essere una caratteristica tipica del PDAC. Conclusioni. La nostra strategia ha identificato marcatori biologici capaci di distinguere differenti comportamenti clinici del tumore in termini di progressione e sede di recidiva. La differente espressione di questi predittori potrebbe essere la causa di differenze biologiche che hanno un effetto sui meccanismi di progressione e diffusione tumorale. Al fine di confermare i nostri risultati stiamo realizzando un tissue microarray di resezioni chirurgiche di pazienti affetti da adenocarcinoma duttale. Inoltre in futuro il modello statistico sviluppato potrà essere applicato per testare ogni nuovo potenziale marcatore; caratterizzata la sua espressione sulle linee cellulari, si procederà con l’analisi inter-linea per verificare se sia un potenziale indicatore di aggressività in vitro o in vivo nel modello murino; quindi si potrà confermare il suo reale ruolo diagnostico, prognostico o predittivo in ambito clinico.

Il carcinoma a cellule squamose è il più frequente tumore maligno del cavo orale (1, 2) ed è la sesta causa di mortalità legata a tumore nel mondo. Esso è caratterizzato da un basso tasso di sopravvivenza, dovuto principalmente alla diagnosi tardiva ed all’elevata frequenza di recidive e/o metastasi. Nel mio studio si è pertanto cercato di individuare dei marcatori a livello salivare che potessero facilitare la diagnosi precoce, utilizzando la metodica SELDI, e verificare le già note potenzialità della Survivina come marcatore biologico, al fine di rendere quindi possibile, in futuro, l’impiego della saliva come mezzo diagnostico, sia sfruttando i vantaggi che la caratterizzano, quali semplicità e rapidità di raccolta, assenza di rischi per l’operatore e minimo disagio per il paziente al momento del prelievo ,sia cercando di risolvere le problematiche legate al suo utilizzo in ambito clinico e non solo laboratoristico. Essendo riusciti nell’intento di evidenziare un nuovo marcatore, auspichiamo che nei prossimi anni si possano continuare le ricerche in modo tale da poterne evidenziare altri e standardizzarne le variazioni in condizioni patologiche.

Background: MPLC represents a diagnostic challenge. Topic of the discussion is how to distinguish these patients as a metastatic or a multifocal disease. While in case of the different histology there are less doubt on the opposite in case of same histology is mandatory to investigate on other clinical features to rule out this question. Matherials and Methods: A retrospective review identified all patients treated surgically for a presumed diagnosis of SPLC. Pre-operative staging was obtained with Total CT scan and fluoro-deoxy positron emission tomography and mediastinoscopy. Patients with nodes interest or extra-thoracic location were excluded from this study. Epidermal growth factor receptor (EGFR) expression with complete immunohistochemical analisis was evaluated. Survival was estimated using Kaplan-Meyer method, and clinical features were estimated using a long-rank test or Cox proportional hazards model for categorical and continuous variable, respectively. Results: According to American College Chest Physician, 18 patients underwent to surgical resection for a diagnosis of MPLC. Of these, 8 patients had 3 or more nodules while 10 patients had less than 3 nodules. Pathologic examination demonstrated that 13/18(70%) of patients with multiple histological types was Adenocarcinoma, 2/18(10%) Squamous carcinoma, 2/18(10%) large cell carcinoma and 1/18(5%) Adenosquamosu carcinoma. Expression of EGFR has been evaluated in all nodules: in 7 patients of 18 (38%) the percentage of expression of each nodule resulted different. Conclusions: MPLC represent a multifocal disease where interactions of clinical informations with biological studies reinforce the diagnosis. EGFR could contribute to differentiate the nodules. However, further researches are necessary to validate this hypothesis.

Background: MPLC represents a diagnostic challenge. Topic of the discussion is how to distinguish these patients as a metastatic or a multifocal disease. While in case of the different histology there are less doubt on the opposite in case of same histology is mandatory to investigate on other clinical features to rule out this question. Matherials and Methods: A retrospective review identified all patients treated surgically for a presumed diagnosis of SPLC. Pre-operative staging was obtained with Total CT scan and fluoro-deoxy positron emission tomography and mediastinoscopy. Patients with nodes interest or extra-thoracic location were excluded from this study. Epidermal growth factor receptor (EGFR) expression with complete immunohistochemical analisis was evaluated. Survival was estimated using Kaplan-Meyer method, and clinical features were estimated using a long-rank test or Cox proportional hazards model for categorical and continuous variable, respectively. Results: According to American College Chest Physician, 18 patients underwent to surgical resection for a diagnosis of MPLC. Of these, 8 patients had 3 or more nodules while 10 patients had less than 3 nodules. Pathologic examination demonstrated that 13/18(70%) of patients with multiple histological types was Adenocarcinoma, 2/18(10%) Squamous carcinoma, 2/18(10%) large cell carcinoma and 1/18(5%) Adenosquamosu carcinoma. Expression of EGFR has been evaluated in all nodules: in 7 patients of 18 (38%) the percentage of expression of each nodule resulted different. Conclusions: MPLC represent a multifocal disease where interactions of clinical informations with biological studies reinforce the diagnosis. EGFR could contribute to differentiate the nodules. However, further researches are necessary to validate this hypothesis.

In recent years, the main end point of immunosuppressive therapy after liver transplantation has moved from the prevention of acute cellular rejection (ACR) toward the preservation of long-term graft function and prevention of immunosuppression-related side effects. This approach requires an optimal management of immunosuppressive therapy according to patient risk factors. However, the concentration of immunosuppressive drugs in the serum of patients, which is generally used as a surrogate for the level of immunosuppression, does not provide information about the magnitude of suppression of the immune system. Therefore a reliable marker for the development of ACR, or to predict patients who could tolerate reduced immunosuppression, would be crucial for improving post-transplant management of liver transplanted patients. The aims of the studies presented in this thesis were: 1) to assess the incidence of ACR after liver transplantation, to identify potential risk factors for ACR, and to evaluate the impact of ACR and its histological severity on outcomes; 2) to evaluate the role of liver function tests and blood eosinophil count as potential biomarkers for ACR after liver transplantation, with special attention on prediction of histologically proven moderate and severe ACR; 3) to evaluate the expression of specific immunological markers for ACR in patients before and after liver transplantation. The results of the studies showed that patient and graft survival at 1, 5 and 10 years after liver transplantation were not different with respect to presence or absence of ACR. Only untreated moderate/severe ACR was associated with increased death/graft loss using adjusted Cox regression analysis, whereas mild ACR, whether treated or not, had no effect. With regards to the evaluation of potential markers of ACR, despite peripheral eosinophilia was not sufficiently predictive of moderate/severe ACR, the delta in eosinophil count between the first and second biopsies was the only independent predictor of histological improvement, irrespective of whether bolus steroids were used. Lastly, we demonstrated that the increased expression of C28 and C38 on both CD4+ and CD8+ T cells and the increased levels of IL-17. These alterations of immune system could be used routinely in clinical practice to assess the immune status of liver transplanted patients and to properly manage immunosuppressive therapy
Lo scopo principale della terapia immunosoppressiva dopo trapianto di fegato è passato dalla prevenzione del rigetto acuto alla preservazione della funzionalità a lungo termine dell’organo trapiantato e alla prevenzione degli effetti collaterali dovuti alla terapia immunosoppressiva. Per perseguire tale scopo è necessaria una gestione ottimale della terapia immunosoppresiva stessa. Tuttavia, la misurazione dei livelli ematici dei farmaci immunosoppressori, generalmente utilizzati come surrogato dei livelli di immunosoppressione, non fornisce informazioni relative alla reale intensità della soppressione del sistema immunitario. Pertanto l’individuazione di marcatori biologici di rigetto acuto e/o di tolleranza risulta fondamentale per poter migliorare la gestione della terapia immunosoppressiva dopo-trapianto di fegato. Gli scopi degli studi riportati in questa tesi sono: 1) determinare l’incidenza e gli eventuali fattori di rischio di rigetto acuto dopo trapianto di fegato, valutare in che l’influenza del rigetto acuto e della sua severità istologica sulla sopravvivenza dell’organo e del paziente dopo trapianto di fegato; 2) valutare il ruolo degli indici di funzionalità epatica e della conta eosinofilica ematica come potenziali marcatori biologici di rigetto acuto dopo trapianto di fegato, in particolare di grado moderato/severo; 3) valutare, prima e dopo trapianto di fegato l’espressione di specifici marcatori immunologici di rigetto acuto. I risultati degli studi condotti hanno evidenziato come pazienti con diagnosi di rigetto acuto alla biopsia di protocollo presentino una sopravvivenza di organo e paziente, a 1, 5 e 10 anni dal trapianto di fegato, del tutto sovrapponibile a quella di pazienti senza evidenza istologica di rigetto acuto alla biopsia di protocollo. L’insorgenza di rigetto acuto di grado moderato/severo non sottoposto a trattamento farmacologico è tuttavia associata ad aumentata incidenza di decesso o perdita dell’organo post-trapianto. Nel valutare potenziali marcatori biologici di rigetto acuto, abbiamo dimostrato che nonostante la conta eosinofilica periferica non sia sufficientemente predittiva per lo sviluppo di rigetto acuto post-trapianto, la differenza nella conta eosinofilica tra la prima e la seconda biopsia epatica può essere considerato un fattore predittivo di miglioramento istologico, indipendentemente dall’utilizzo o meno di terapia con boli steroidei. Non è stata invece evidenziata alcuna associazione tra l’alterazione degli indici di funzionalità epatica e l’insorgenza di rigetto acuto. Infine, è stato dimostrato che l’insorgenza di rigetto acuto risulta associata ad aumentata espressione di CD28 e CD38 sia sui linfociti T CD4+ che CD8+ e ad un aumento dei livelli di IL-17. Tali alterazioni del sistema immunitario potrebbero essere utilizzate nella pratica clinica per valutare lo stato di soppressione del sistema immunitario in pazienti sottoposti a trapianto di fegato con il fine ultimo di una gestione ottimale e personalizzata della terapia immunooppressiva

Introduzione: I soggetti con disturbi mentali hanno un rischio più elevato di comportamenti suicidari rispetto alla popolazione generale. Pertanto, ad oggi, i ricercatori hanno studiato alcuni marcatori biologici potenzialmente correlati al suicidio. Nonostante molti studi abbiano riportato una possibile correlazione tra bassi livelli ematici di lipidi e tentativo di suicidio, sono emersi risultati contrastanti. Scopo del lavoro: Abbiamo studiato se il colesterolo totale , il colesterolo LDL e i trigliceridi ematici siano associati a recenti tentativi di suicidio in soggetti con diversi disturbi mentali. Metodi: Abbiamo condotto uno studio trasversale su 593 pazienti ricoverati di volta in volta, affetti da disturbo dello spettro di schizofrenico, disturbo bipolare, depressivo maggiore e di personalità. I livelli ematici dei lipidi sono stati confrontati tra soggetti ricoverati per un recente tentativo di suicidio e quelli senza tale storia recente. Inoltre, secondo l'ipotesi che collega impulsività e violenza con bassi livelli ematici di lipidi, è stata valutata l'associazione tra i livelli lipidici e il tentativo di suicidio con metodo violento. Risultati: Non abbiamo trovato alcuna associazione di colesterolo totale e LDL e trigliceridi con i tentativi di suicidio, analizzando anche per diagnosi e metodo di suicidio. Inoltre, un'analisi statistica post-hoc ha mostrato una tendenza verso la significatività (p = 0,06) nell'associazione tra un alto livello di colesterolo (≥160 mg / dL) e il recente tentativo di suicidio. Conclusioni: I nostri risultati non supportano l'ipotesi di associazione tra profili lipidici e tentativi di suicidio in soggetti con diversi disturbi mentali. Sono necessarie ulteriori ricerche per chiarire il ruolo dei marcatori biologici nei comportamenti suicidari.
Background: Subjects with mental disorders have a higher risk of suicide behaviors than the general population. So, to date, researchers have investigated some biomarkers possibly related to suicidality. Despite many studies have reported a possible relationship between low lipid serum levels and suicide attempt, conflicting results have emerged. Aim We investigated whether serum total cholesterol, LDL cholesterol and triglycerides are associated with recent suicide attempts in subjects with different mental disorders. Methods We conducted a cross-sectional study, including 593 consecutively admitted inpatients with schizophrenia spectrum, bipolar, major depressive, and personality disorders. Serum lipid levels were compared between subjects admitted for a recent suicide attempt and those without such recent history. Moreover, according to hypothesis that links impulsivity and violence with low serum lipid levels, the association between lipid levels and violent suicide attempt was assessed. Results We did not find any association of total and LDL cholesterol and triglycerides with suicide attempts, also considering diagnosis and suicide methods. In addition, a post-hoc analysis showed a trend toward significance (p=0.06) in the association between high cholesterol level (≥160mg/dL) and recent suicide attempt. Conclusions Our results do not support the hypothesis of association between lipid profiles and suicide attempts in subjects with different mental disorders. Further research is needed to clarify the role of biomarkers in suicidal behaviors.

Purpose: to quantify the mRNA levels of MMP-3, MMP-9, VEGF and Survivin in peripheral blood and the serum levels of CA-125, Ca19-9 in women with and without endometriosis and to investigate the performance of these markers to differentiate between deep and ovarian endometriosis. Methods: a case controls study enrolled a series of 60 patients. Twenty controls have been matched with 20 cases of ovarian and 20 cases of deep endometriosis. Univariable and multivariable performance of serum CA125 and CA19-9, mRNA for Survivin, MMP9, MMP3 and VEGF genes have been evaluated by means of ROC curves and logistic regression respectively. Results: No difference in markers concentration were detected between ovarian and deep endometriosis. In comparison with controls serum CA19 and CA125 yielded the better sensitivity followed by mRNA for Survivin gene (81.5%, 51.9% and 7.5% at 10% false positive rate respectively). Multivariable estimated odds of endometriosis yielded a sensitivity of 87% at the same false positive rate. Conclusions: A combination of serum and molecular markers could allow a better diagnosis of endometriosis.

Purpose: to quantify the mRNA levels of MMP-3, MMP-9, VEGF and Survivin in peripheral blood and the serum levels of CA-125, Ca19-9 in women with and without endometriosis and to investigate the performance of these markers to differentiate between deep and ovarian endometriosis. Methods: a case controls study enrolled a series of 60 patients. Twenty controls have been matched with 20 cases of ovarian and 20 cases of deep endometriosis. Univariable and multivariable performance of serum CA125 and CA19-9, mRNA for Survivin, MMP9, MMP3 and VEGF genes have been evaluated by means of ROC curves and logistic regression respectively. Results: No difference in markers concentration were detected between ovarian and deep endometriosis. In comparison with controls serum CA19 and CA125 yielded the better sensitivity followed by mRNA for Survivin gene (81.5%, 51.9% and 7.5% at 10% false positive rate respectively). Multivariable estimated odds of endometriosis yielded a sensitivity of 87% at the same false positive rate. Conclusions: A combination of serum and molecular markers could allow a better diagnosis of endometriosis.

This chapter provides an overview of the current computational methods for PPI network cleansing. The authors first present the issue of identifying reliable PPIs from noisy and incomplete experimental data. Next, they address the questions of which are the expected results of the different experimental studies, of what can be defined as true interactions, of which kind of data are to be integrated in assigning reliability levels to PPIs and which gold standard should the authors use in training and testing PPI filtering methods. Finally, Marcatili and Tramontano describe the state of the art in the field, presenting the different classes of algorithms and comparing their results. The aim of the chapter is to guide the reader in the choice of the most convenient methods, experiments and integrative data and to underline the most common biases and errors to obtain a portrait of PINs which is not only reliable but as well able to correctly retrieve the biological information contained in such data.